Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.

OBJECTIVETo evaluate the feasibility of genetic and prenatal diagnosis for a family affected with pyruvate kinase deficiency (PKD).

METHODSTargeted sequence capture and high-throughput sequencing technology was used to detect the exons and exon-intron boundaries of the PKLR gene in a clinically suspected PKD patient. Meanwhile, the genotype of the pedigree was validated by Sanger sequencing. Prenatal genetic diagnosis was performed by amniotic fluid sampling after genotype of the mother of the proband was determined.

RESULTSThe proband was found to harbor double heterozygous mutations, c.661G>A (Asp221Asn) and c.1528C>T (Arg510Ter), which resulted in amino acid substitution Asp221Asn and Arg510Ter. Such mutations were confirmed by Sanger sequencing. The mother and father of the proband were detected to have respectively carried c.1528C>T (Arg510Ter) and c.661G>A (Asp221Asn) mutation. The fetus was found to have carried the same mutations as the proband. Following selected abortion, analysis of fetal tissue was consistent with the result of prenatal diagnosis.

CONCLUSIONThe compound mutations of c.661G>A and c.1528C>T of PKLR gene probably underlie the PKD in the family. Prenatal diagnosis of the mutations analysis can facilitate detection of affected fetus in time.

Adult ; Anemia, Hemolytic, Congenital Nonspherocytic ; embryology ; enzymology ; genetics ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Pyruvate Kinase ; deficiency ; genetics ; metabolism ; Pyruvate Metabolism, Inborn Errors ; embryology ; enzymology ; genetics

Objective To explore the relationship between heart rate variability (HRV) and deceleration capacity (DC) in children with idiopathic ventricular premature contraction of different origins. Methods The clinical data from 155 children with idiopathic ventricular premature contraction were retrospectively analyzed. According to the age, the children were divided into young children group (

OBJECTIVETo investigate the therapeutic effect of heating and bandage treatment for chronic lymphedema of extremities accompanied with erysipelas.

METHODSFrom March 2004 to March 2013, 80 patients with chronic lymphedema of extremities accompanied with erysipelas were analyzed retrospectively. The patients underwent heating treatment (42 degree centigrade) with infrared light machine made by Shanghai Ninth People's Hospital, 2 hours a day, 20 hours for a session. Bandage treatment was adopted after heating treatment. 1 or 2 sessions were performed for each patient every year. The erysipelas occurring frequency, patients subjective feeling, treatment sessions and elastic material usage was recorded during the follow-up period. The erysipelas occurring frequency was tested by the method of rank and inspection. SPSS 17. 0 was used for statistical analysis.

RESULTSAfter heating and bandage treatment, the occurrence frequency of erysipelas was obviously controlled (Z = 7.598, P = 0.000). Erysipelas was not occurred any more in 60 (75%)patients. Remarkable reduction of occurrence frequency of erysipelas caused by various reasons was showed after treatment. Primary and secondary lymphedema after treatment were compared with those before treatment respectively, showing statistical difference (Z = 3.417 and 5.009, P = 0.001 and 0.000). Most of patients felt better subjectively. The relapse rate of erysipelas and lymphedema was lower if keeping using elastic material to give more pressure on extremities after therapy.

CONLUSIONSHeating and bandage treatment can obviously reduce the occurrence frequency of erysipelas. It can improve the quality of patients' lives. Simultaneously, the subsequent elastic material pressure therapy is essential.

Bandages ; Chronic Disease ; Combined Modality Therapy ; methods ; Erysipelas ; complications ; therapy ; Extremities ; Female ; Humans ; Hyperthermia, Induced ; methods ; Lymphedema ; complications ; therapy ; Middle Aged ; Pressure ; Recurrence ; Retrospective Studies ; Time Factors

Objective To understand the clinical distribution and monitoring the change of resistance of Streptococcus pneumoni‐ae ,effective for clinical anti infection to provide reference .Methods Using VITEK 2 Compact to analyze the bacteria identification and drug sensitivity data ,and using WHONET5 .3 software and SPSS13 .0 software for statistical analysis .Results From 2008 to 2013 ,588 strains of Streptococcus pneumoniae were isolated ,mainly distributed in the intensive care unit (ICU) ,followed by respir‐atory department of internal medicine ,general pediatrics ;mainly from sputum samples ,followed by the throat swabs and blood samples .The highest resistant rate was erythromycin ,followed by penicillin and cotrimoxazole ;Streptococcus pneumoniae remains sensitive to ofloxacin ,levofloxacin ,vancomycin ,linezolid ,chloramphenicol .Conclusion The resistance rate of Streptococcus pneu‐moniae was rising ,and that great attention should be paid to the bacterial drug resistance so as to reasonably use a antibiotics based on the result of drug susceptibility testing .

Objectives To explore the clinical application of deceleration capacity of rate (DC), acceleration capacity of rate (AC) and heart rate variability (HRV) in children with precardial distress of unknown origin. Methods A total of 56 children with precardial distress of unknown origin and 63 healthy children aged 6 to 17 years were examined by 24 h dynamic elec-trocardiogram, and the indexes of DC and HRV were compared between these two groups. Results DC value of children with precardial distress is less than that of the control group (P<0.05), AC value is greater than that of the control group (P<0.05), and heat rate (HR) is greater than that of the control group (P<0.05). No statistical differences were observed in the indexes of HRV between the two groups. The indexes of DC show a signiifcant positive correlation with HRV in children with precardial distress(r=0.27~0.40, P<0.05), while appear a negative relation with HR (r=-0.46, P=0.000). In contrast, the indexes of AC show a signiifcant negative correlation with HRV (r=-0.57~-0.34, P<0.05), and appears a positive relation with HR(r=0.61, P=0.000). HR value is higher in male children less than 12 years old with precardial distress than that of age-matched males in control group, and RMSSD is lower than the latter. DC value of male children more than 12 years with precardial distress is lower than that of age-matched males in control group, while AC value is higher than that of the latter;DC value is signiifcant lower in fe-male children more than 12 yeares with precardial distress than that of age-matched females in the control group (P<0.05). Con-clusions The activity of vagus nerve in children with precardial distress of unknown origin is decreased. DC value is signiifcantly lower than that of control group, and shows correlation with indexes of HRV. There is no signiifcant difference in DC and HRV value between male and female children with precardial distress. DC value is lower in children aged 12 or older with precardial distress than that of age-matched children in the control group, which indicates adolescents are vulnerable to autonomic nerve functional disorder.

Objective To investigate the effects of 2-methoxyestradiol( 2-ME) on apoptosis of K562 cells and its mechanisms. Methods The K562 cells were cultured and divided into three groups. The control group: K562 cells were cultured without 2-ME treatment. The experimental group: K562 cells were cultured in the medium containing different concentrations of 2-ME (1, 2, 4, 8 and 16 μmol/L) for 36 h. The negative control group: K562 cells were replaced by water without RNase in the medium containing different concentrations of 2-ME for 36 h. The apoptosis rate, the protein and its mRNA expression of Caspase-3 and XIAP, the activity of superoxide dismutase (SOD) and reactive oxygen species (ROS) of K562 cells wasdetected by TUNEL, flow cytometry (FCM), half-quantitative RT-PCR and xanthenes oxidized enzyme assay,respectively. Results After treated with 2-ME at different concentrations for 36 hours, in the specified concentration range, 2-ME induced apoptosis of K562 cells in a concentration-dependent manner. The possible functional mechanism of 2-ME was to up-regulate Caspase-3 but down-regulate XIAP mRNA expression, and increase ROS activity but decrease SOD activity. Conclusion 2-ME can induce apoptosis of K562 cells in a concentration-dependent manner and indicate its promising potential in the treatment of chronic myeloid leukemia(CML) patients.

The gene smu. 776 encodes a possible S-adenosylmethionine-dependent methyltransferase of 385 residues in Streptococcus mutans, a primary pathogen for human dental caries. The DNA fragment of smu.776 was cloned into pET28a and expressed in good amount from the E. coli strain BL21 (DE3). Smu.776 protein was purified to homogeneity in a two-step procedure ofNi2+ chelating and size exclusion chromatography. Crystals were obtained by hanging-drop vapor diffusion method and diffracted to 2.0 (A) resolution.The crystal belongs to orthorhombic space group C2 with cell dimension of a=168.47 (A), b= 50.66 (A), c=53.96 (A), β=104.22°. The asymmetric unit is expected to contain one molecule with solvent content of 51.3%.

Smu. 260 encodes a putative protein of 200 residues in Streptococcus mutans, a primary pathogen for human dental caries. Smu. 260 was cloned into expression vector pET28a and expressed in good amount trom the E. coli strain BL21 (DE3). Smu.260 protein was purified to homogeneity in a two-step procedure of Ni2+ chelating and size exclusion chromatography. The purified protein exists in two forms, a dimer form about 46 ku with yellow color and a tetramer form without apparent color. Crystals were obtained from the dimer protein by hanging-drop vapor-diffusion method. The crystals diffracted to about 2.3 A resolution and belong to orthorhombic space group P212121 with cell dimensions of a = 89.88A, b = 90.91 A, c = 105.17 A. The asymmetric unit is expected to contain two dimers with solvent content of 53%.

Objective To investigate the clinical features, etiological classification and staging of epiretinal macular membrane(MEM). Methods Clinical materials of 194 cases of MEM diagnosed by fundus fluorescein angiography in outpatient department of eye clinic in this hospital from 1983 to 2000 were retrospectively analyzed. Results There were typical clinical symptoms and signs of MEM in all of this 222 eyes of 194 patients. Etiological classification revealed that 4 cases were congenital(2.12%), 22 cases were secondary(11.34%), and 168 cases were idiopathic(86.60%). Staging of course of disease indicated that 119 eyes were in early stage(53.60%), 72 eyes were in middle stage(32.43%), and 31 eyes were in late stage(13.96%). Conclusion MEM may be classified as congenital, secondary and idiopathic type according to its pathogenesis, as early, middle and late stage according to the clinical course of disease. This can be helpful in treating the disease.